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Systematical targeted multicomponent characterization and comparison of Arnebiae Radix and its three confusing species by offline two-dimensional liquid chromatography/LTQ-Orbitrap mass spectrometry

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Abstract

Arnebiae Radix, commonly known as “Zicao,” can be easily confused with other compounding species, posing challenges for its clinical use. Here, we developed a comprehensive strategy to systematically characterize the diverse components across Arnebiae Radix and its three confusing species. First, an offline two-dimensional liquid chromatography (2D-LC) system integrating hydrophilic interaction chromatography (HILIC) and reverse phase (RP) separations was established, enabling effective separation and detection of more trace constituents. Second, a polygonal mass defect filtering (MDF) workflow was implemented to screen target ions and generate a precursor ion list (PIL) to guide multistage mass (MSn) data acquisition. Third, a three-step characterization strategy utilizing diagnostic ions and neutral losses was developed for rapid determination of molecular formulas, structure classes, and compound identification. This approach enabled systematic characterization of Arnebiae Radix and its three confusing species, with 437 components characterized including 112 shikonins, 22 shikonfurans, 144 phenolic acids, 131 glycosides, 18 flavonoids, and 10 other compounds. Additionally, 361, 230, 340, and 328 components were identified from RZC, YZC, DZC, and ZZC, respectively, with 142 common components and 30 characteristic components that may serve as potential markers for distinguishing the four species. In summary, this is the first comprehensive characterization and comparison of the phytochemical profiles of Arnebiae Radix and its three confusing species, advancing our understanding of this herbal medicine for quality control.

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Funding

The work was supported by Shanghai Sailing Program (NO. 21YF1455800), Qi-Huang Chief Scientist Project of National Administration of Traditional Chinese Medicine (2020), and Science and Technology Major Project of Inner Mongolia (No. 2021ZD0017).

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  1. Fei Sha and Jianqing Zhang contributed equally to this work.

Authors and Affiliations

  1. School of Pharmacy, Hangzhou Normal University, Zhejiang, 311121, Hangzhou, China

    Fei Sha & Dean Guo

  2. Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Science, Haike Road 501, Shanghai, 201203, China

    Jianqing Zhang, Wenlong Wei, Cuicui Wang, Xiaolan Li, Xuanjing Shen, Yaling An, Jiayuan Li & Dean Guo

  3. School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, People’s Republic of China

    Huanya Yang & Yunshu Hu

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  1. Fei Sha
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Sha, F., Zhang, J., Yang, H. et al. Systematical targeted multicomponent characterization and comparison of Arnebiae Radix and its three confusing species by offline two-dimensional liquid chromatography/LTQ-Orbitrap mass spectrometry. Anal Bioanal Chem 416, 583–595 (2024). https://doi.org/10.1007/s00216-023-05067-x

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